G. Steigleder
Oct 1, 1960
Citations
0
Influential Citations
2
Citations
Journal
Journal of Investigative Dermatology
Abstract
In previous reports we have described a simple method for determining the activity and observing the behavior of certain enzymes (mainly nonspecific esterases) on the skin surface (6-8). For this purpose the azo-dye coupling reaction of Nachlas and Seligman, as modified by Gomori (1), was performed in a glass chamber placed directly on the skin surface (Figure 1). The incubation medium consisted of 50 mg diazoblue B and 50 mg 1-naphthylacetate dissolved in a few drops of acetone and diluted in 50 ce Sorensen phosphate buffer, pH 7.4. In these studies each glass chamber contained 2 cc of this solution. When esterases are present, the 1-naphthylacetate is hydrolyzed to 1-naphthol and acetic acid. The naphthol couples with the diazo blue and a clearly visible azodye is formed, which then stains the surface of the skin. Furthermore, it is also possible to determine quantitatively the amount of 1-naphthol or acetic acid formed and how much of the azodye is developed (9). Our previous studies have shown that the intensity of skin color is proportional to the activity of non-specific esterases on the skin surface. It was also found that covering the surface of the skin with white petrolatum prior to incubation, blocks the reaction completely (5).